桑树MnERF2的表达分析  

Expression Analysis of MnERF2 in Mulberry

作　　者：董亚茹[1] 赵东晓[1] 耿兵 李云芝[1] 王照红[1] DONG Ya-ru;ZHAO Dong-xiao;GENG Bing;LI Yun-zhi;WANG Zhao-hong(The Sericulture Research Institute of Shandong Province,Yantai 264002)

机构地区：[1]山东省蚕业研究所,烟台264002

出　　处：《生物技术通报》2022年第11期112-121,共10页Biotechnology Bulletin

基　　金：山东省自然科学基金(ZR2019PC043);烟台市科技创新发展计划(2021XDHZ057);山东省现代农业产业技术体系蚕桑产业创新团队(SDAIT-18-03);国家蚕桑产业技术体系烟台综合试验站(CARS-18-SYZ08)。

摘　　要：乙烯响应因子(ethylene responsive factors,ERFs)是植物特异性转录因子,参与多种生物过程,特别是非生物胁迫耐受。研究干旱胁迫下MnERF2的表达情况,为研究桑树AP2/ERF转录因子的抗逆分子机制提供基础理论数据。从桑树中鉴定了一个ERF转录因子亚家族成员,命名为MnERF2,对其进行生物信息学分析,并通过瞬时转化分别获得过表达MnERF2和RNA干扰(RNAi)沉默MnERF2桑树植株,利用功能增益和功能损失方法分析其抗旱功能。结果显示,MnERF2开放阅读框为1 050 bp,编码349个氨基酸,编码蛋白质分子量为39.48 kD,理论等电点为5.6,具有一个典型的AP2/ERF保守结构域。过表达MnERF2显著提高桑树对干旱胁迫的耐受性,相比之下,MnERF2的沉默显著降低了对干旱胁迫的耐受性。进一步试验表明,MnERF2增强SOD、POD、CAT、GST活性,促进ASA、GSH、脯氨酸含量的升高,降低O_(2)^(·-)、H_(2)O_(2)、·OH、MDA的含量和电解质渗透率。MnERF2通过提高脯氨酸的生物合成来调节渗透势,通过提高抗氧化酶活性和抗氧化物质含量来提高活性氧的清除能力,从而提高桑树干旱胁迫的耐受性。Ethylene responsive factors(ERFs)is a plant-specific transcription factor that involves in various biological processes,especially abiotic stress tolerance.The expression of MnERF2 under drought stress was studied to provide basic theoretical data for understanding the stress resistance molecular mechanism of AP2/ERF transcription factor in mulberry.A member of the ERF transcription factor subfamily,named MnERF2,was identified from mulberry,and analyzed by bioinformatics.Transiently transfected mulberry plants with transiently overexpressed MnERF2 and RNAi-silenced MnERF2 were generated for its drought resistance via gain-and loss-of-function analysis.The results showed that the open reading frame of MnERF2 was 1 050 bp,encoding 349 amino acids.The relative molecular weight of MnERF2 protein was 39.48 kD,with isoelectric point 5.6 and a typical AP2/ERF conserved domain.Overexpression of MnERF2 significantly elevated the tolerance of mulberry to drought stress.In contrast,silencing of MnERF2 significantly decreased the tolerance to drought stress.Further experiments demonstrated that MnERF2 enhanced the activities of SOD,POD,CAT and GST,increased the contents of ASA,GSH and proline,while decreased the contents of O_(2)^(·-),H_(2)O_(2),·OH,MDA and electrolyte permeability.MnERF2 regulated osmotic potential by increasing proline contents,and enhanced the ability of ROS scavenging by increasing antioxidant enzyme activity and antioxidant substance content,thus improved drought stress tolerance in mulberry.

关 键 词：桑树 AP2/ERF转录因子 干旱胁迫 瞬时表达 

分 类 号：S888.2[农业科学—特种经济动物饲养]